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Differential induction of chemokines in gut epithelial cells by bacterial and viral infection
A886
AGA ABSTRACTS
• PSEUDOCHOLINESTERASE ISOZYMES DURING ACUTE PHASE IN CROHN'S DISEASE. G., Novacek, W. Reinisch, H. Vogelsang, S. Kapiotis, B. Gmeiner. Department of Internal Medicine IV, Gastroenterology and Hepatology, University of Vienna, Austda. The percentage distribution of the pseudocholinesterase (PCHE) isozymes (C1, C2, C3 and C4) is significantly altered in patients with active Crohn's disease (CD). The aim of the present Study was to assess the time course and the clinical valence of the PCHE isozymes during an acute phase in CD. Ten healthy controls and 10 patients with active CD (CDAI>200) were examined. In patients with CD an acute phase treatment with prednisolone was administered for 7 weeks with 6 patients reaching remission (CDAI<150). PCHE isozymes were detected by electrophoretic separation. Results: Before initiation of steroid treatment the isozymes C1 and C4 were decreased and increased, respectively, and did not change dudng the time of acute phase treatment (Table). There was no difference between controls and patients with CD in the isozymes C2 and C3 (Table). And there was no difference between patients who reached remission and who did not in all isezymes at the end of prednisolone therapy. Five of the patients were followed up for the. subsequent 3 months of remission. Dudng this pedod C1 and C4 normalized (T lble). Controls CD (n=10), CD (n=10). CD (n=5), Start of End of 3 months of (n=10) prednisolone prednisolone remission 279(248-304) 134(80-245) # 77 (60-112) # CDAI 2.1 (1.1-4.9)* 1.2 (1-1.6) *# CRP(mg/dl) 0.4 (0.3-0.4) 5.3 (2.7-7)* PCHE(kU/I) 4.6 (4.1-5.5) 3.4 (2.8-3,7)* 4.1 (3.5-4.9) 4.8 (4.5-5.7) C1 (%) 20 (16-22) 6 (5-8)* 9 (8-10)* 18 (15-20) # 8 (6-11) 9 (8-11) 16 (15-17) *# C2 (%) 11 (10-12) C3 (%) 6 (5-7) 5 (3-9) 7 (5-10) 8 (7-9) c4/%) 54/5g-71) 82 (73-85)* 76 (70-79)* 59 (58-61)# Median (interquartile range); *p<0.01 vs controls; #p<0.05 vs start of prednisolone; CRP = C-reactive protein. Conclusion: The percentage distribution of the PCHE isozymes remains altered dudng acute phase treatment in CD. Normalisation reflects the therapeutical result of complete remission during the following months. Therefore, the PCHE isozymes are useful parameters in CD to indicate active CD or long term remission.
• E X P R E S S I O N OF V A S C U L A R A D D R E S S I N S IN I N F L A M M A T O R Y B O W E L DISEASE. J. O'Brien*, A. Chitale*, P. Streeter, M. Donovan; *Division of G a s t r o e n t e r o l o g y , W a s h i n g t o n U n i v e r s i t y School of M e d i c i n e and S e a r l e R e s e a r c h Department, St. Louis, Missouri. V a s c u l a r A d d r e s s i n s (VA) are a group of a d h e s i o n molecules which are expressed on endothelial venules. V A can be induced on e n d o t h e l i u m at sites of c h r o n i c inflammation but their role in Inflammatory Bowel Disease (IBD) is yet to be determined. Therefore, w e c o n d u c t e d a blinded, p r o s p e c t i v e study to d e t e r m i n e the p r e s e n c e of V A in IBD p a t i e n t s (pts) [ U l c e r a t i v e Colitis ( U . C . ) - 7 , Crohn's (C.D.) - 9] and c o n t r o l s - 9. A clinical IBD a c t i v i t y score ( H a r v e y - B r a d s h a w Score) and a 4 grade endoscopic score w a s d e t e r m i n e d for e a c h patient. B i o p s i e s (Bx) w e r e o b t a i n e d from the m o s t prominently i n f l a m e d mucosa. A MECA-79 immunoc h e m i s t r y stain w a s p e r f o r m e d on each intestinal Bx. This IgG a n t i b o d y stain d e t e c t e d the V A p r o t e i n a n t i g e n s a s s o c i a t e d w i t h venules. In a d d i t i o n a DAB (peroxidase stain) was also o b t a i n e d for a qualitative m e a s u r e m e n t of n e u t r o p h i l s present. F o l l o w i n g DAB s t a i n i n g a c h l o r o e s t e r a s e stain was a l s o p e r formed. The p r e s e n c e of V A is a q u a l i t a t i v e finding; h o w e v e r an a r b i t r a r y g r a d i n g s y s t e m r a n g i n g from no V A p r e s e n t to 3+ w a s used. E x p r e s s i o n of V A in the c o l o n i c BX w a s i n t e r p r e t e d in a b l i n d e d fashion. Results: All U.C. p t s e x c e p t one h a d p o s i t i v e MECA-79 s t a i n i n g i n d i c a t i n g t h e p r e s e n c e of VA. However, all C.D. pts (except one) and all control pts h a d n e g a t i v e M E C A - 7 9 staining, E x c e p t for one U.C. pt all IBD pts h a d a p o s i t i v e p e r o x i d a s e stain d e n o t i n g the p r e s e n c e of PMNS. The c h l o r o e s t e r a s e stain a l s o t e n d e d to be p o s i t i v e in all pts. Conclusion: T h e r e w a s e x p r e s s i o n of V A on intestinal Bx p r e s e n t in U.C. p t s b u t not C.D. pts. PMN's w e r e p r e s e n t in all IBD pts. The s i g n i f i c a n c e of a p o s i t i v e c h l o r o e s t e r a s e stain is uncertain. VA may act as a m a r k e r of d i s e a s e a c t i v i t y and m a y p o s s i b l y p l a y a r o l e in the d e v e l o p m e n t of new therapies.
GASTROENTEROLOGY, Vol. 1 0 8 , No. 4
• ASSOCIATION OF ULCERATIVE COLITIS AND ISCHEMIC HEART DISEASE. H. Nuutinen, A. Reunanen, M. F~irkkil/i, K. Sepp~il~i, T.A. Miettinen. Second Department of Medicine, University of Helsinki, Research and Development Centre, The Social Insurance Institution, Helsinki, Finland The age specific prevalence of ulcerative colitis (UC) have been shown to have a bimodal age distribution with peaks identified in the 30---39 and greater than or equal to 60-year age. Because the second peak is in the same age-range than the peak for ischemic heart disease, and because inflammatory bowel disease (IBD) patients have been suggested to represent a high risk group with regard to thromboembolic events, we decided to study if there is any association between ischemic heart disease and UC. The prevalence figures were calculated from the records of the Social Insurance Institution of Finland, which can give us reliable information about the prevalence of these diseases among the 5 million population of Finland. The prevalence of coronary heart disease in the older age group of patients with UC (>60 years) was significantly higher both in males (p<.001)(M) and in females (p<.05)(F) than in the general population (see below). As far as we know this is the first report of the association of IBD and ischemic heart disease. Although the reason for this striking increase of ischemic heart disease in patients with UC is not known, at least two possible factors may contribute. First the hypercoagulability in IBD could increase the risk for ischemic heart disease. Secondly patients with ischemic heart disease may also have ischemic bowel disease which could be one pathogenetic factor in inflammatory bowel disease especially in the older people. PREVALENCEOF UC IN PREVALENCEOF CORONARYHEART FINLANDIN 1990 DISEASEIN 1990 0
300 ]
M(uc)
20
AGE-GROUP(YEARSI
~ uc)
AGE-GROUP(YEARS)
• DIFFERENTIAL INDUCTION OF CHEMOKINES IN GUT EPITHELIAL CELLS BY BACTERIAL AND VIRAL INFECTION. M. Ode, P.B. Ernst, B. Prabhakar, S. Patibandla, G. Klimpel, R. Garofalo, P.L. Ogra, S.E. Crowe. Depts. of Internal Medicine, Pediatrics, and Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX. Recent studies indicate that invasive enteric bacteria increase the expression of IL-8 in intestinal epithelial cell lines and that non-invasiva H, py[or/ induces IL-8 in cultured gastric epithelial cells. That the epithelium may play a role in the recruitment and activation of immune cells via the release of chemokines is important to our understanding of the pathogenesis of enteric diseases. However, less is known about GI epithelial expression of other chemokines or their regulation by viral infection. We Compared constitutive and stimulated mRNA expression of IL-8, MIP-IG, MCP-1, and RANTES by PCR and the presence of immunoreactiva protein for IL-8, MIP-la and RANTES by ELISA in four human GI epithelial cell lines (T84, HT-29, Caco-2, and Kato III) in response to. PMA, cytokines, and following exposure to Salmonella typhimurium, poliovirus, and respiratory syncytial virus (RSV). IL-8 mRNA and protein was detected in all cell lines tested under basal conditions and both were stimulated by PMA and Salmonella infection. Constitutive and PMA- or Salmonella-stimulated MCP-1 and MIP-la were detected by PCR in T84 and Caco-2 but not HT-29 or Kato II1. RANTES was expressed in 3 of 4 lines as measured by PCR but protein levels measured in cell supernates were low (except in T84). Although Salmonella infection was a potent inducer of immunoreactive IL-8, we were unable to detect any effect of Salmonella on the induction of immunoreactive RANTES or MIP-la. Poliovirus infection of HT-29 and Caco-2 cells was associated with cytopathic effect (CPE) and viral replication but no increased expression of IL-8, RANTES or MIP-la. In contrast, poliovirus did not replicate or induce CPE to the same degree in T84 as in HT-29 and Caco-2 cells. However, inoculation of T84 with either poliovirus or RSV increased immunoreactive IL-8, RANTES and MIP-la. These results suggest that gastrointestinal epithelial cells express a broad range of chemokines which are differentially regulated by bacteria and viruses. These patterns of chemokine production may partially account for the different cellular infiltrate observed with viral and bacterial pathogens. Supported by AGA Foundation/IndUstry Research Scholar Award and the CCFA.
AGA ABSTRACTS
• PSEUDOCHOLINESTERASE ISOZYMES DURING ACUTE PHASE IN CROHN'S DISEASE. G., Novacek, W. Reinisch, H. Vogelsang, S. Kapiotis, B. Gmeiner. Department of Internal Medicine IV, Gastroenterology and Hepatology, University of Vienna, Austda. The percentage distribution of the pseudocholinesterase (PCHE) isozymes (C1, C2, C3 and C4) is significantly altered in patients with active Crohn's disease (CD). The aim of the present Study was to assess the time course and the clinical valence of the PCHE isozymes during an acute phase in CD. Ten healthy controls and 10 patients with active CD (CDAI>200) were examined. In patients with CD an acute phase treatment with prednisolone was administered for 7 weeks with 6 patients reaching remission (CDAI<150). PCHE isozymes were detected by electrophoretic separation. Results: Before initiation of steroid treatment the isozymes C1 and C4 were decreased and increased, respectively, and did not change dudng the time of acute phase treatment (Table). There was no difference between controls and patients with CD in the isozymes C2 and C3 (Table). And there was no difference between patients who reached remission and who did not in all isezymes at the end of prednisolone therapy. Five of the patients were followed up for the. subsequent 3 months of remission. Dudng this pedod C1 and C4 normalized (T lble). Controls CD (n=10), CD (n=10). CD (n=5), Start of End of 3 months of (n=10) prednisolone prednisolone remission 279(248-304) 134(80-245) # 77 (60-112) # CDAI 2.1 (1.1-4.9)* 1.2 (1-1.6) *# CRP(mg/dl) 0.4 (0.3-0.4) 5.3 (2.7-7)* PCHE(kU/I) 4.6 (4.1-5.5) 3.4 (2.8-3,7)* 4.1 (3.5-4.9) 4.8 (4.5-5.7) C1 (%) 20 (16-22) 6 (5-8)* 9 (8-10)* 18 (15-20) # 8 (6-11) 9 (8-11) 16 (15-17) *# C2 (%) 11 (10-12) C3 (%) 6 (5-7) 5 (3-9) 7 (5-10) 8 (7-9) c4/%) 54/5g-71) 82 (73-85)* 76 (70-79)* 59 (58-61)# Median (interquartile range); *p<0.01 vs controls; #p<0.05 vs start of prednisolone; CRP = C-reactive protein. Conclusion: The percentage distribution of the PCHE isozymes remains altered dudng acute phase treatment in CD. Normalisation reflects the therapeutical result of complete remission during the following months. Therefore, the PCHE isozymes are useful parameters in CD to indicate active CD or long term remission.
• E X P R E S S I O N OF V A S C U L A R A D D R E S S I N S IN I N F L A M M A T O R Y B O W E L DISEASE. J. O'Brien*, A. Chitale*, P. Streeter, M. Donovan; *Division of G a s t r o e n t e r o l o g y , W a s h i n g t o n U n i v e r s i t y School of M e d i c i n e and S e a r l e R e s e a r c h Department, St. Louis, Missouri. V a s c u l a r A d d r e s s i n s (VA) are a group of a d h e s i o n molecules which are expressed on endothelial venules. V A can be induced on e n d o t h e l i u m at sites of c h r o n i c inflammation but their role in Inflammatory Bowel Disease (IBD) is yet to be determined. Therefore, w e c o n d u c t e d a blinded, p r o s p e c t i v e study to d e t e r m i n e the p r e s e n c e of V A in IBD p a t i e n t s (pts) [ U l c e r a t i v e Colitis ( U . C . ) - 7 , Crohn's (C.D.) - 9] and c o n t r o l s - 9. A clinical IBD a c t i v i t y score ( H a r v e y - B r a d s h a w Score) and a 4 grade endoscopic score w a s d e t e r m i n e d for e a c h patient. B i o p s i e s (Bx) w e r e o b t a i n e d from the m o s t prominently i n f l a m e d mucosa. A MECA-79 immunoc h e m i s t r y stain w a s p e r f o r m e d on each intestinal Bx. This IgG a n t i b o d y stain d e t e c t e d the V A p r o t e i n a n t i g e n s a s s o c i a t e d w i t h venules. In a d d i t i o n a DAB (peroxidase stain) was also o b t a i n e d for a qualitative m e a s u r e m e n t of n e u t r o p h i l s present. F o l l o w i n g DAB s t a i n i n g a c h l o r o e s t e r a s e stain was a l s o p e r formed. The p r e s e n c e of V A is a q u a l i t a t i v e finding; h o w e v e r an a r b i t r a r y g r a d i n g s y s t e m r a n g i n g from no V A p r e s e n t to 3+ w a s used. E x p r e s s i o n of V A in the c o l o n i c BX w a s i n t e r p r e t e d in a b l i n d e d fashion. Results: All U.C. p t s e x c e p t one h a d p o s i t i v e MECA-79 s t a i n i n g i n d i c a t i n g t h e p r e s e n c e of VA. However, all C.D. pts (except one) and all control pts h a d n e g a t i v e M E C A - 7 9 staining, E x c e p t for one U.C. pt all IBD pts h a d a p o s i t i v e p e r o x i d a s e stain d e n o t i n g the p r e s e n c e of PMNS. The c h l o r o e s t e r a s e stain a l s o t e n d e d to be p o s i t i v e in all pts. Conclusion: T h e r e w a s e x p r e s s i o n of V A on intestinal Bx p r e s e n t in U.C. p t s b u t not C.D. pts. PMN's w e r e p r e s e n t in all IBD pts. The s i g n i f i c a n c e of a p o s i t i v e c h l o r o e s t e r a s e stain is uncertain. VA may act as a m a r k e r of d i s e a s e a c t i v i t y and m a y p o s s i b l y p l a y a r o l e in the d e v e l o p m e n t of new therapies.
GASTROENTEROLOGY, Vol. 1 0 8 , No. 4
• ASSOCIATION OF ULCERATIVE COLITIS AND ISCHEMIC HEART DISEASE. H. Nuutinen, A. Reunanen, M. F~irkkil/i, K. Sepp~il~i, T.A. Miettinen. Second Department of Medicine, University of Helsinki, Research and Development Centre, The Social Insurance Institution, Helsinki, Finland The age specific prevalence of ulcerative colitis (UC) have been shown to have a bimodal age distribution with peaks identified in the 30---39 and greater than or equal to 60-year age. Because the second peak is in the same age-range than the peak for ischemic heart disease, and because inflammatory bowel disease (IBD) patients have been suggested to represent a high risk group with regard to thromboembolic events, we decided to study if there is any association between ischemic heart disease and UC. The prevalence figures were calculated from the records of the Social Insurance Institution of Finland, which can give us reliable information about the prevalence of these diseases among the 5 million population of Finland. The prevalence of coronary heart disease in the older age group of patients with UC (>60 years) was significantly higher both in males (p<.001)(M) and in females (p<.05)(F) than in the general population (see below). As far as we know this is the first report of the association of IBD and ischemic heart disease. Although the reason for this striking increase of ischemic heart disease in patients with UC is not known, at least two possible factors may contribute. First the hypercoagulability in IBD could increase the risk for ischemic heart disease. Secondly patients with ischemic heart disease may also have ischemic bowel disease which could be one pathogenetic factor in inflammatory bowel disease especially in the older people. PREVALENCEOF UC IN PREVALENCEOF CORONARYHEART FINLANDIN 1990 DISEASEIN 1990 0
300 ]
M(uc)
20
AGE-GROUP(YEARSI
~ uc)
AGE-GROUP(YEARS)
• DIFFERENTIAL INDUCTION OF CHEMOKINES IN GUT EPITHELIAL CELLS BY BACTERIAL AND VIRAL INFECTION. M. Ode, P.B. Ernst, B. Prabhakar, S. Patibandla, G. Klimpel, R. Garofalo, P.L. Ogra, S.E. Crowe. Depts. of Internal Medicine, Pediatrics, and Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX. Recent studies indicate that invasive enteric bacteria increase the expression of IL-8 in intestinal epithelial cell lines and that non-invasiva H, py[or/ induces IL-8 in cultured gastric epithelial cells. That the epithelium may play a role in the recruitment and activation of immune cells via the release of chemokines is important to our understanding of the pathogenesis of enteric diseases. However, less is known about GI epithelial expression of other chemokines or their regulation by viral infection. We Compared constitutive and stimulated mRNA expression of IL-8, MIP-IG, MCP-1, and RANTES by PCR and the presence of immunoreactiva protein for IL-8, MIP-la and RANTES by ELISA in four human GI epithelial cell lines (T84, HT-29, Caco-2, and Kato III) in response to. PMA, cytokines, and following exposure to Salmonella typhimurium, poliovirus, and respiratory syncytial virus (RSV). IL-8 mRNA and protein was detected in all cell lines tested under basal conditions and both were stimulated by PMA and Salmonella infection. Constitutive and PMA- or Salmonella-stimulated MCP-1 and MIP-la were detected by PCR in T84 and Caco-2 but not HT-29 or Kato II1. RANTES was expressed in 3 of 4 lines as measured by PCR but protein levels measured in cell supernates were low (except in T84). Although Salmonella infection was a potent inducer of immunoreactive IL-8, we were unable to detect any effect of Salmonella on the induction of immunoreactive RANTES or MIP-la. Poliovirus infection of HT-29 and Caco-2 cells was associated with cytopathic effect (CPE) and viral replication but no increased expression of IL-8, RANTES or MIP-la. In contrast, poliovirus did not replicate or induce CPE to the same degree in T84 as in HT-29 and Caco-2 cells. However, inoculation of T84 with either poliovirus or RSV increased immunoreactive IL-8, RANTES and MIP-la. These results suggest that gastrointestinal epithelial cells express a broad range of chemokines which are differentially regulated by bacteria and viruses. These patterns of chemokine production may partially account for the different cellular infiltrate observed with viral and bacterial pathogens. Supported by AGA Foundation/IndUstry Research Scholar Award and the CCFA.